Inactivation of the polyanionic detergent sodium polyanetholsulfonate by hemoglobin

Sodium polyanetholsulfonate (SPS) has been added to blood culture media for many years. Its incorporation results in a higher yield of positive blood cultures due to its inactivation of antimicrobial cationic compounds. The most active of these cations include complement components, aminoglycoside-aminocyclitol antibiotics, and receptors on polymorphonuclear leukocytes. There have been reports from studies conducted outside patient blood culture bottles that SPS itself may possess antibacterial activity against some isolates of Neisseria meningitidis, Neisseria gonorrhoeae, and Peptostreptococcus anaerobius. Conversely, in patient clinical trials there has been no significant difference in pathogen isolation rates in the presence or absence of SPS. In an attempt to explain this in vitro/in vivo disparity, a search was undertaken to elucidate which variable constituent in blood, heretofore not studied quantitatively, might have a major effect on modulating the activity of SPS. It was found that hemoglobin combined stoichiometrically with SPS with a Kd of approximately 10(-7) mol/liter. Optimum SPS inactivation occurred at an SPS/hemoglobin ratio of 1:6 (wt/wt). SPS-sensitive isolates of N. gonorrhoeae and N. meningitidis were protected by the addition of hemoglobin from the antimicrobial effects of this polyanion in time-kill studies. This protection was directly related to the amount of SPS combined in solution. Therefore, the amount of free hemoglobin in solution must be measured when studying the antimicrobial activity of polyanions or when evaluating the effect of different polyanions on the recovery rates of pathogens in patient blood culture clinical trials.